Murine il 1β

LN and spleen were isolated by dissection from mice and then mashed through a 70-µm filter. Spleen cells were lysed in ammonium-chloride-potassium lysis buffer to remove red blood cells. Cells isolated from spleen and LNs were counted, and 1-5 × 10 6 cells were first stained in PBS and Ghost Live/Dead (Tonbo), followed by blocking in 2.4G2 before staining with the appropriate antibodies for flow cytometry. For transcription factor staining, cells were fixed overnight in the eBioscience Foxp3/Transcription Factor/Fixation-Concentrate kit (Thermo Fisher Scientific). After fixation, cells were permeabilized and stained with the appropriate antibodies. For ICS, cells were stimulated for 4 h in Brefeldin A (eBioscience) and eBioscience Cell Stimulation Cocktail (500×). Cells were then fixed and permeabilized using the BD cytofix/cytoperm kit before staining with the appropriate antibodies.
Islets were purified following standard collagenase protocols as described 22 and dissociated by incubating with a non-enzymatic solution (Sigma, St, Lois, MO) followed by trituration per the manufacturer's instructions.
were purchased from R&D Systems (murine IL-4, murine IL-6, and human IL-2), Miltenyi (murine IL-12, murine IL-1β, and murine IL-23), or HumanKine (human TGFβ).